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1996 Fiscal Year Final Research Report Summary

Gene expression control of animal lectins containing a collagen-like domain.

Research Project

Project/Area Number 07672354
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biological pharmacy
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

KAWASAKI Nobuko  College of Medical Tecnology, Kyoto University Professor, 医療技術短期大学部, 教授 (70077676)

Co-Investigator(Kenkyū-buntansha) KAWASAKI Toshisuke  Pharmaceutcal Sciences Kyoto University Professor, 薬学部, 教授 (50025706)
Project Period (FY) 1995 – 1996
KeywordsConglutinin / Animal lectin / Collagen-like structure / Bovine serum / Expression control of gene / Luciferase / Promoter / Transcription regulatory sequence
Research Abstract

Conglutinin is a unique bovine plasma protein which mediates the agglutination of the sensitized erythrocyte-solid phase iC3b (conglutination). The protein is identified as a Ca^<2+>-dependent animal lectin specific for N-acetylglucosamine and a member of proteins which have a collagen-like domain (collectins, collagen-like lectins).
In this research, the 5'-regulatory region sequence and the expression control of conglutinin gene were studied to elucidate the biological function of conglutinin with respect to the gene structure.
1) Characterization of 5'-upstream region of the conglutinin gene.
The 5'-flanking region of the conglutinin gene was cloned and sequenced by gene walking using vector (cassette) -ligation mediated PCR.In the promoter region (700bp) of the gene, several putative consensus sequences that may be involved in the expression of conglutinin were identified.
2) Analysis of the transcription regulatory sequences with the reporter gene assay.
A 5'-flanking region of the con … More glutinin gene (700bp) was examined for its promoter activity using luciferase as a reporter gene. To determine the region of the conglitinin promoter responsible for the transcription, a series of 5'-stepwise promoter deletions and site-directed mutations were constructed into the luciferase plasmids. These plamid-promoter DNA constructs were transfected into the human hepatoma cell line HepG2. The luciferase promoter assays revealed that an unknown positively controlling cis-acting elements exists around-200bp. This unknown cis-element acted synergistically with the AP-1 sequence present at-158bp--154bp.
3) Gel-shift assay and DNase I footprint analysis of the transcription regulatory sites.
The bindings of the unknown cis-acting element (around-200bp) and AP-1 (-158--154bp) with the nuclear protein extracts of the HepG2 cell and bovine liver cells were demonstrated with gelmobility shift assays. The DNase I footprint analysis identified the binding sites of the unknown cis-acting element with a trans-acting element in the nuclear protein around-180bp. Less

  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] N. Kawasaki: "Gene structure and expression control of bovine conglutinin : A C-type mammalian lectin containing a collagen-like domain." Glycoconjugate Journal. 12-4. 550 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T. Kawasaki: "New aspects on the role of mannan-binding protein (MBP) and conglutinin." Immunology. 86-Suppl. 1. 2 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 上村和秀: "生体防御に関与するカルシウム依存性動物レクチンの構造と機能の相関" 実験医学. 13-18. 46-51 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] N.Kawasaki: "Gene structure and expression control of bovine conglutinin : A C-type mammalian lectin containing a collagen-like domain." Glycoconjugate Journal. 12-4. 550 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Kawasaki: "New aspects on the role of mannan-binding protein (MBP) and conglutinin." Immunology. 86-Suppl.1. 2 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Uemura: "Structure-function relationships in the calcium-dependent animal lectins." Experimental Medicine (in Japanese). 13-18. 46-51 (1995)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-03-09  

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